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The TRANSCEND trial1 studied a cohort of more than 250 patients with relapsed/refractory non-Hodgkin's lymphoma treated with the chimeric antigen receptor T-cell (CART) product, lisocabtagene maraleucel. This trial enrolled a high number of patients and included a relatively long follow-up (36 months). Estimating the gain in overall survival (OS) for CART patients versus non-CART controls suffers from the lack of long-term survival information for CART. One advantage of the TRANSCEND trial is the availability of survival results at 36 months; this allows us to reliably determine the survival gain. To carry out this estimation, we kept in mind that the restricted mean survival time (RMST) is recognised to be the best outcome measure for managing long-term survival …

Aims We applied the restricted mean survival time (RMST) to analyse the survival data reported in the PARADIGM‐HT trial in which sacubitril + valsartan was studied in comparison with enalapril in patients with heart failure. The estimates of this parameter were compared with the published values of hazard ratio (HR). Methods Two endpoints were evaluated: a composite of death or hospitalization and cardiovascular death. Our analyses were performed by considering the original follow‐up of 41.4 months and on the basis of a lifetime perspective. All statistical calculations were carried out using specific packages developed under the R‐platform. Results According to our RMST analysis, the results for the composite endpoint in the comparison of sacubitril + valsartan vs. enalapril showed an improvement from 32.9 to 34.2 months (gain of 1.25 months). This result is based on a time horizon of 41.4 months. The results for the cardiovascular mortality endpoint showed a RMST of 37.2 months for sacubitril + valsartan vs. 36.2 for enalapril (gain of 0.96 months). In the two lifetime analyses, the improvements were much more relevant and yielded a gain of 25.8 months for the composite endpoint and 27.6 months for survival free from cardiovascular death. Conclusions Using the data of the PARADIGM‐HT trial, our analysis confirmed that the RMST has documented advantages over the HR, particularly when the clinical study is characterized by a long follow‐up. The number needed to treat (NNT) has a more specific methodological role and cannot be replaced by the RMST.

Background: The purpose of this study was to assess the effectiveness of immune checkpoint inhibitors (ICIs) in advanced urothelial carcinoma. Materials & methods: We selected seven cohorts of patients published in four clinical trials. The restricted mean survival time (RMST) was used to analyze survival curves, perform the comparisons and rank the treatments based on their effectiveness. The performance of RMST was compared with that of a network meta-analysis. Results: Three ICIs, vinflunine and best standard care, given as second line, were examined. ICIs significantly improved overall survival compared with best standard care. However, the survival gain was quite limited (up to 2.27 months). Post hoc pairwise comparisons were calculated. Conclusion: Our results summarized the efficacy of these treatments and confirmed the good methodological performance of RMST.

NMR metabolomics represents a powerful tool to characterize the cellular effects of drugs and gain detailed insight into their mode of action. Here, we have exploited NMR metabolomics to illustrate the changes in the metabolic profile of A2780 ovarian cancer cells elicited by auranofin (AF), a clinically approved gold drug now repurposed as an anticancer agent. An early and large increase in intracellular glutathione is highlighted as the main effect of the treatment accompanied by small but significant changes in the levels of a few additional metabolites; the general implications of these findings are discussed in the frame of the current mechanistic knowledge of AF.

International audience; The reactions of the medicinal gold(i) compound auranofin and its close analogues with vasopressin and the diselenide analogue were comparatively investigated by LC-electrospray MS/MS. Evidence is gained of the possible cleavage of the S-S and Se-Se bridges induced by Au(i). Notably, we found that, in the absence of reducing agents, the sulfur and selenium atoms are metallated only at high temperature (70 °C) with the preferential binding of gold to selenium. The reaction with the S-S bridge can take place at physiological temperature (37 °C) under reducing conditions. The implications of these results are discussed in the general frame of the reactivity of biologically relevant soft Lewis acids with peptides and proteins.

Three novel platinum(II) complexes bearing N-heterocyclic ligands, i.e., Pt2c, Pt-IV and Pt-VIII, were previously prepared and characterized. They manifested promising in vitro anticancer properties associated with non-conventional modes of action. To gain further mechanistic insight, we have explored here the reactions of these Pt compounds with a few model proteins, i.e., hen egg white lysozyme (HEWL), bovine pancreatic ribonuclease (RNase A), horse heart cytochrome c (Cyt-c) and human serum albumin (HSA), primarily through ESI MS analysis. Characteristic and variegate patterns of reactivity were highlighted in the various cases that appear to depend both on the nature of the Pt complex and of the interacting protein. The protein-bound Pt fragments were identified. In the case of the complex Pt2c, the adducts formed upon reaction with HEWL and RNase A were further characterized by solving the respective crystal structures: this allowed us to determine the exact location of the various Pt binding sites. The implications of the obtained results are discussed in relation to the possible mechanisms of action of these innovative anticancer Pt complexes.

Arsenoplatins are adducts of two chemically important anticancer drugs, cisplatin and arsenic trioxide, that have a Pt(II) bond to an As(III) hydroxide center. Screens of the NCI-60 human tumor cell lines reveal that arsenoplatin-1 (AP1), [Pt(μ-NHC(CH(3))O)(2)ClAs(OH)(2)], the first representative of this novel class of anti-cancer agents, displays a superior activity profile relative to the parent drugs As(2)O(3) or cisplatin in majority of cancer cell lines tested. These activity profiles are important because the success of arsenic trioxide in blood cancers (such as APL) has not been seen in solid tumors due to the rapid clearance of arsenous acid from the body. To understand the biological chemistry of these compounds, we evaluated interactions of AP-1 with the two important classes of biomolecules – proteins and DNA. The first structural studies of AP-1 bound to model proteins reveal that platinum(II) binds the Nε of His in a manner that preserves the Pt-As bond. We find that AP-1 readily enters cells and binds to DNA with an intact Pt-As bond (Pt:As ratio of 1). At longer incubation times, however, the Pt:As ratio in DNA samples increases, suggesting that the Pt-As bond breaks and releases the As(OH)(2) moiety. We conclude that arsenoplatin-1 has the potential to deliver both Pt and As species to a variety of hematological and solid cancers.

Spinal muscular atrophy (SMA) is a rare neuromuscular disorder. Currently there are two approved drug treatments for SMA: nusinersen and onasemnogene abeparvovec (OAX). The purpose of the present s...

Lichen sclerosus (LS) is a chronic relapsing disease with a predilection for the anogenital areas of patients of both sexes and at all ages. In males, LS is usually a late diagnosis, increasing the...

Very limited information exists from Italian hospitals about the utilization and expenditure of medical devices. The ratio expenditure/DRG reimbursement (expressed as a percentage) is a useful parameter, particularly when it is separately calculated for each individual Azienda and, within each Azienda, according to the medical discipline. To generate benchmarks in this area, we have made reference to the expenditure incurred for medical devices from January to June 2019 in all hospitals of the Tuscany region. These expenditures divided by medical discipline have been compared with the DRG reimbursements that hospitals receive from our Region for the same disciplines. This benchmark is represented by percent ratio expenditure/reimbursement for each of the eight Aziende and, within each Azienda, according to the medical specialty. These percentages indicate, for each medical discipline, to what extent medical devices typically consume the reimbursement resulting from the DRGs. These benchmarks facilitate the interpretation of values estimated locally. Determinants leading to ‘excessive’ values (e.g. >100%) of this parameter can include an extensive use of costly devices, a high frequency of non-remunerative procedures, the presence of inappropriate treatments, the need to update specific DRGs, deficiencies in coding expenditures and procedures, and so on.

A few gold compounds were recently found to show antimicrobial properties in vitro, holding great promise for the discovery of new drugs to overcome antibiotic resistance. Here, the inhibition of the bacterial virulence factor urease by four Au(I)-compounds, namely Au(PEt3)Cl, Au(PEt3)Br, Au(PEt3)I and [Au(PEt3)2]Cl, obtained from the antiarthritic Au(I)-drug Auranofin and earlier reported to act as antimicrobials, is investigated. The three monophosphino Au(I) complexes showed IC50 values in the 30–100 nM range, while the diphosphino Au(I) complex, though being less active, still showed a IC50 value of 7 μM. The structural basis for this inhibition was provided by solving the crystal structures of urease co-crystallized with Au(PEt3)I and [Au(PEt3)2]Cl: at least two Au(I) ions bind the enzyme in a flap domain involved in the catalysis, thus obliterating enzyme activity. Peculiar changes observed in the two structures reveal implications for the mechanism of soft metal binding and enzyme inactivation.

Introduction: Extra-mammary Paget’s disease is a rare form of intraepithelial adenocarcinoma with a variable pattern of invasion. Vulvar Paget’s disease represents 1% of all vulvar cancers. Standard treatment is surgical excision, however, the recurrence rate is high and surgery leads to anatomical, functional and sexual morbidity. Some studies suggest the efficacy of photodynamic therapy following CO2 Laser irradiation as treatment for this disease. Methods: In the present study, ten patients with vulvar Paget’s disease underwent fractional carbon dioxide (CO2) laser abrasion, followed by 3 h of occlusive application of aminolaevulinic acid (ALA) and then 100 J/cm2 irradiation with a 630-nm lamp. This combination treatment regime was repeated every 2 weeks for a total of 5 times. Results: At the end of the 12-month follow-up period, only two patients maintained a complete remission of the disease and recurrence was observed in all patients who had originally shown a partial remission. Conclusions: According to our experience, Photodynamic Therapy-ALA following fractional CO2 Laser does not seem to be an effective treatment for vulvar EMPD in the long term.

Background Scarce is knowledge on the process regulating the development of acid secretion, orexigenic signaling, and chemosensing in the stomach of young pigs. Changes of early microbial encounters by suckling pigs can interact with the gut maturation, by the induction of different molecular signaling. Our goal was to assess if the age of offspring and the maternal environment, influenced by sow antibiotic treatment peripartum, could affect gastric morphology and the expression of genes involved in the control of hydrochloric secretion, feed intake, taste, and inflammation in offspring stomach. Methods 84 pigs from sows fed a diet with amoxicillin (on –d10 to +d21 from farrowing, ANT) or without (CON) were sacrificed at d14, d21, d28 (weaning) or d42. Samples of oxyntic (OXY), pyloric (PY) and cardiac mucosae close to OXY were collected and parietal and enteroendocrine cells (EECs) were counted. Relative gene expression of a set of 11 key genes (ATP4A, SSTR2, GAST, GHRL, MBOAT4, PCSK1, GNAT1, TAS1R1, TAS1R3, IL8 and TNF) was assessed by qRT-PCR. In addition, 40 offspring obtained from the same ANT and CON sows were offered a normal or a fat-enriched diet for 4 weeks between 140 and 169 d of age, and then OXY and PY were sampled. Results The number of parietal and EECs increased with age (P ATP4A increased with age (within suckling, P = 0.043, post-weaning vs. suckling, P P GHRL increased during suckling (P = 0.012), and post-weaning as a trend (P = 0.088). MBOAT4 tended to increase during suckling (P = 0.062). TAS1R1 increased from suckling to post-weaning period (P =0.001) and was lower in ANT offspring (P = 0.013). GNAT1 in PY was higher in ANT offspring (P = 0.041). Antibiotic treatment of sows peripartum increased expression of GHRL and MBOAT4 in OXY of growing-finishing offspring aged 5 months. Conclusions Data show that sensing for umami taste and ghrelin regulation can be affected by maternal environment, but the development of acid secretion, orexigenic signaling and taste perception in the stomach are mostly developmentally controlled.

Background Maintenance therapy using poly (ADP-ribose) polymerase inhibitors (PARPIs) is an important therapeutic option in advanced ovarian cancer after platinum-based chemotherapy. Materials and methods We evaluated randomized studies (n = 5) describing the effect of maintenance therapy with PARPIs; they were obtained mainly by searching PubMed. Patient data for the analysis were derived from progression-free survival curves. Restricted mean survival time (RMST) and 95% confidence interval were estimated for individual arms of each trial. Results The three PARPIs used (olaparib, niraparib, rucaparib) all showed a higher effectiveness than placebo. The gains in progression-free survival were 6 - 8 months. Conclusion Maintenance therapy studies provide evidence that olaparib, niraparib, rucaparib are effective treatments for advanced ovarian cancer.

Programmed cell death ligand 1 (PD-L1) and programmed cell death protein 1 (PD-1) inhibitors are increasingly used in a variety of solid tumors. In patients with DNA mismatch repair-deficient (dMMR)/microsatellite instability-high (MSI-H) metastatic colorectal cancer, their efficacy has been demonstrated in recently published phase-II trials. However, an indirect comparison of effectiveness between pembrolizumab, nivolumab, and nivolumab+ipilimumab is not yet available. After a standard literature search, we analyzed four overall survival (OS) curves from three phase-II trials. Individual patient data were reconstructed from each curve using a specific web-based technique (Shiny method). Indirect statistical comparisons were made based on hazard ratio (HR) and restricted mean survival time (RMST). Nivolumab+ipilumumab had a better HR compared with pembrolizumab (0.65, 95% confidence interval [CI], 0.43 to 1.002, p=0.051); the difference being close to statistical significance. In the analysis based on RMST, the combination of nivolumab+ipilimumab showed a significantly longer OS than pembrolizumab (improvement in RMST, 1.08 mos; 95%CI, 0.11 to 2.06; p=0.029). The other two pairwise differences in RMST (nivolumab vs. pembrolizumab and nivolumab+ ipilimumab vs. nivolumab) had a smaller magnitude (0.25 mos, 95%CI, -0.99 to 1.48, and 0.84 mos, 95%CI, -0.40 to 2.07, respectively) and were far from statistical significance. Our results favoring the combination of nivolumab+ipilimumab in metastatic colorectal cancer must be viewed with caution owing to the indirect nature of our statistical comparisons. With this limitation in mind, the magnitude of the incremental benefit for the above combination treatment was estimated to be around one month over a follow-up of 15 months.

Introduction In conducting a survival meta-analysis, the typical methodological approach analyses the hazard ratios (HRs) of individual trials and then combines them into a pooled meta-analytical estimate. The length of follow-up of individual trials is not generally accounted for. Recent techniques aimed at individual patient-data reconstruction from Kaplan-Meier graphs represent an important methodological innovation. These techniques permit the combination of the survival curves published in a single clinical trial but are also applicable to more than one trial. In the case of multiple trials, a meta-analysis can be conducted without using any statistical model of meta-analysis. Methods As an example of this new approach, we applied a technique of individual patient data reconstruction to the Kaplan-Meier graphs of overall survival reported in two phase-III trials, which were conducted on patients with locally advanced/advanced non-small cell lung cancer selected according to their PD-L1 expression status, not previously treated for their metastatic disease. Only subjects with PD-L1 ≥50% were considered for our analysis. The experimental arms received pembrolizumab monotherapy while the control arms were given platinum-based chemotherapy. The survival graphs were obtained for both trials. For each Kaplan-Meier curve, the graph was firstly digitalized. Then, the Shiny package was used to reconstruct patient-level data. Finally, the pooled survival curves were generated from the reconstructed patient-level data along with the relevant Cox statistics; for this purpose, we used three packages (“coxph”, “survfit”, and “ggsurvplot”) under the R-platform. Results In our pooled analysis based on this procedure, we compared 453 patients given pembrolizumab vs. 451 controls given chemotherapy. The HR estimated from reconstructed patient-level data was 0.670 (95% confidence interval [CI], 0.566 to 0.793). Conclusion The analysis described herein demonstrates the easy applicability of the Shiny technique. This technique was successful in generating a pooled survival graph for the experimental treatment groups vs. controls and efficiently estimated the pooled HR in which the results of the two trials were combined.

A grape pomace extract (GPext) and a grape seed extract (T) have been melt mixed within poly(butylene succinate) (PBS) and tested as natural stabilizers. Their effect on the PBS stabilization has been evaluated trough thermo-mechanical degradative tests (reprocessing), thermo-oxidation (oven aging), thermogravimetric analysis (TGA) and biodegradation tests. GPext and T have been deeply analyzed in terms of polyphenolic profile and bioactivity to connect the stabilization's results with the antioxidant's structures. Additive T exhibited the best results, maintaining unaltered the PBS molecular weight both after six reprocessing steps and after 300 h of oven aging and to increase the degradation temperature of more than 20 °C. These encouraging results have been explained by the long-chains polyphenols present within T (medium degree of polymerization of 10.8), by the excellent radical scavenger activity of T (6.0 mM TEAC) and by the low releases of peroxides from T (0.06 µM H2O2).

The current COVID pandemic crisis made it even clearer that the solutions to several questions that public health must face require the access to good quality data. Several issues of the value and potential of health data and the current critical issues that hinder access are discussed in this paper. In particular, the paper (i) focuses on “real-world data” definition; (ii) proposes a review of the real-world data availability in our country; (iii) discusses its potential, with particular focus on the possibility of improving knowledge on the quality of care provided by the health system; (iv) emphasizes that the availability of data alone is not sufficient to increase our knowledge, underlining the need that innovative analysis methods (e.g., artificial intelligence techniques) must be framed in the paradigm of clinical research; and (v) addresses some ethical issues related to their use. The proposal is to realize an alliance between organizations interested in promoting research aimed at collecting scientifically solid evidence to support the clinical governance of public health.

Background Lichen sclerosus is an autoimmune dermatosis that in women typically involves vulvar area. This condition can strongly impact on the quality of life.To date, topical steroids are the most effective treatment, although adverse effects are possible, especially in long-term application. The aim of this study is to investigate the efficacy of fractional CO2 laser in reducing symptoms of vulvar lichen sclerosus (VLS) during maintenance therapy with topical steroids (application twice weekly or less);(ii) to assess how long this reduction of symptoms persists during followup; (iii) histological comparison before and after treatment.To our knowledge, this is the first prospective study evaluating this treatment in women with VLS. Methods Women with a diagnosis of VLS were prospectively enrolled and treated with fractional CO2 laser every 2 months, for a total of 3 sessions.Four questionnaires were periodically administered: the Dermatology Quality of Life Index (DLQI), the Female Sexual Functional Index (FSFI) and 2 specific questionnaires created to assess the severity of disease. Results 23 adult women were enrolled. The fractional CO2 laser treatment significantly improved the scores of all scales from baseline to T4 questionnaires. Conclusions Fractional CO2 laser proved to be effective on VLS symptoms and can be considered an alternative to corticosteroids during maintenance therapy. Larger studies with a control group and randomization are needed to safely generalize our findings.

Hintergrund und ziele In ausgewahlten Fallen ist die konventionelle photodynamische Therapie (cPDT) bei der Behandlung des Basalzellkarzinoms (BZK) eine geeignete Alternative zu einem chirurgischen Eingriff. Allerdings ist sie auf superfizielle BZK beschrankt. Die Vorbehandlung von BZK mit ablativen Lasern konnte die Wirksamkeit der cPDT erhohen. Wir haben die Kombinationstherapie aus cPDT und CO2 -Laser bei der Behandlung von superfiziellen und nodularen BZK untersucht. Patienten und methoden In der prospektiven monozentrischen Interventionsstudie wurden Patienten mit BZK unter Verwendung eines kontinuierlichen supergepulsten CO2 -Lasers fur nodulare BZK sowie eines fraktionierten CO2 -Lasers fur superfizielle BZK mit CO2 -Lasertherapie vorbehandelt. Anschliesend wurden alle Patienten mit photodynamischer Therapie unter Verwendung von Methylaminolavulinat-Creme und einer Aktilite CL128® Lampe (Galderma) behandelt. Ergebnisse 32 Patienten (20 Manner, 12 Frauen) im Alter von 45 bis 96 Jahren (mit insgesamt 181 BZK) erhielten eine Kombinationstherapie aus CO2 -Laser und cPDT. Nach drei Monaten lag die Heilungsrate bei 100 %. Wahrend der durchschnittlichen Nachbeobachtungszeit von 10,7 Monaten (Bereich: 4-18 Monate) gab es bei 97,2 % der Falle keine Anzeichen eines Rezidivs. Wir beobachteten leichte unerwunschte Reaktionen und gute asthetische Ergebnisse. Schlussfolgerungen Aufgrund der hohen Wirksamkeit, dem guten asthetischen Ergebnis und den geringen Nebenwirkungen empfehlen wir diese Kombinationstherapie bei ausgewahlten Fallen.

Background: Given limitations in economic resources, achieving sustainability in healthcare is an increasingly important issue driving policy decisions. Colorectal cancer (CRC) is a frequent oncological diagnosis with an incidence that is expected to increase, making the efficient utilisation of resources a major priority for its diagnosis and treatment. To assess potentially inappropriate utilisation of services in the integrated care pathway of patients with malignant CRC, a series of key performance indicators (KPIs) have been developed by a multidisciplinary panel of the E.Pic.A. project (Economic Appropriateness of an Integrated Care Pathway).Methods: The KPIs identified had to comply with criteria of reliability, representativeness, accessibility and operativity. A definition is provided for each of the KPIs, along with the methodology used to calculate it and a reference target.Results: Eight KPIs were identified that can measure inadequacies of services provided in the diagnostic and treatment pathways for CRC: four for instrumental diagnostics, three for surgery and one for oncologic therapy.Conclusions: Use of the methodology described can help payors to obtain detailed information on inappropriate and wasteful use of healthcare resources, which would then permit their reallocation for interventions with higher value for patients with CRC and other pathologies. (HTA & Market Access, Oncology)

Background: Due to its high incidence, evaluating performance of care delivered to breast cancer patients is a crucial issue. The multidisciplinary panel E.Pic.A. (Economic Appropriateness of an Integrated Care Pathway) defined a set of key performance indicators (KPIs) to evaluate economic waste in breast cancer healthcare interventions. Methods: The E.Pic.A. panel identified the principal KPIs that are crucial within the breast cancer care pathway to evaluate the performance of care. KPIs were defined taking into account their reliability, validity, usability and feasibility of measurement through the linkage between multiple routine healthcare data sources. Results: Seven KPIs were identified: three on instrumental diagnostics, two on surgery and two on treatment. The three KPIs regarding instrumental diagnostics are aimed at assessing the inappropriateness of diagnostic tests performed before and after the index surgery. The two KPIs regarding surgery measure the inappropriateness of possible repeated interventions considering the time elapsed from the index surgery. The two KPIs regarding oncologic therapy measure the inappropriateness about the administration time of adjuvant therapy and radiotherapy considering the time elapsed from the index surgery. Conclusion: E.Pic.A methodology could help to evaluate economic waste in healthcare interventions with the objective of redirecting resources to interventions with greater value. (HTA & Market Access)

Blood glucose concentration control is a classic negative feedback problem with insulin secreted by the pancreas as a control variable. Type 1 Diabetes is a chronic metabolic disease caused by a cellular-mediated autoimmune destruction of the pancreas beta-cells, so exogenous insulin administration is needed to regulate the glycaemia. Postprandial glucose regulation is typically based on the knowledge of an estimation of the ingested carbohydrates, of the Carbohydrate-to-insulin ratio, of the correction factor, of the insulin still active and of a measure of the glycaemia just before the meal. Despite the use of this information meal compensation is yet a key unsolved issue. In this letter a new approach based on machine-learning methodologies is proposed to improve postprandial glucose regulation. The proposed approach uses the multiple K-nearest neighbors classification algorithm to predict postprandial glucose profile due to the nominal therapy and to suggest a correction to time and/or amount of the meal bolus. This approach has been successfully validated on the adult in silico population of the UVA/PADOVA simulator, which has been accepted by the Food and Drug Administration as a substitute to animal trials.

Auranofin (AF) and its iodido analog, i.e., Au(PEt3) I (AFI), were reported to exhibit very promising anticancer properties both in vitro and in vivo. However, both these gold compounds have a scarce aqueous solubility that hampers their pharmaceutical use. Here, we explore whether encapsulation of these metallodrugs inside hydroxypropyl-beta–cyclodextrin (HPβ–CD) may lead to an improved biopharmaceutical profile for the resulting adducts. Phase solubility studies, performed at 25 °C in an aqueous buffer, revealed, in both cases, the formation of a 1:1 drug to cyclodextrin complex; a far greater apparent stability constant (K1:1) was measured for AFI compared to AF (331 M−1 versus ca. 30 M−1). NMR studies conducted on the AFI/HPβ–CD system confirmed the formation of a stable 1:1 adduct. Then, binary systems of AF and AFI with HPβ–CD were prepared by colyophilization and characterized by DSC and PXRD. The results revealed the occurrence of drug complexation and/or amorphization for the AFI/HPβ–CD binary system. Afterwards, the antiproliferative properties of the two cyclodextrin adducts and of the corresponding free drugs were comparatively evaluated in vitro in three representative ovarian cancer cell lines, i.e., A2780, SKOV3, and IGROV-1. The results, in all cases, point out that CD complexation of the two gold drugs does not substantially affect their biological activity. The implications of these findings are discussed in the frame of the current knowledge of AF and its analogs.

Recent accelerating global warming with increasing climate variability exerts a strong impact on terrestrial carbon budgets, but the ecosystem response to the changing climate and the overall climate-vegetation coupling remain largely unclear during different stages of the growing season. The timing of growing seasons can be modulated by different environmental conditions (e.g., thermal and hydrological changes) and affect the overall interpretation of regional climate-vegetation coupling. Here, we analyse the climate-vegetation coupling for Europe during 1982–2014 using a grid-wise definition of the growing season period based on remote sensing data. We quantify sub-seasonal anomalies of vegetation greenness from long-term vegetation indices (Normalized Difference Vegetation Index and two-band Enhanced Vegetation Index), and their relationships with corresponding local growing conditions (2m temperature, downwards surface solar radiation and root-zone soil moisture); and with multiple climate variability indices that reflect the large-scale climatic conditions over Europe. We find that early growing season anomalies in vegetation greenness tend to be large during the first two months of the growing season and that the coupling of these anomalies with large-scale climate largely determines the full-year climate-vegetation coupling. The North Atlantic Oscillation (NAO) and Scandinavian Pattern (SCA) phases evaluated one to two months before the start of growing season are the dominant drivers of the early growing season climate-vegetation coupling over large parts of boreal and temperate Europe. However, the sign of the effect of these indices on vegetation greenness is opposite. The East Atlantic Pattern (EA) evaluated several months in advance of the growing season is instead a main controlling factor on the temperate belt and the Mediterranean region. These findings highlight the importance of accounting for the spatial heterogeneity of growing season periods using location-specific definitions when studying large-scale land-atmosphere interactions.

Chloro(triethylphosphine)gold(I), (Et3PAuCl hereafter), is an Auranofin (AF)-related compound showing very similar biological and pharmacological properties. Like AF, Et3PAuCl exhibits potent antiproliferative properties in vitro toward a variety of cancer cell lines and is a promising anticancer drug candidate. We wondered whether Et3PAuCl encapsulation might lead to an improved pharmacological profile also considering the likely reduction of unwanted side-reactions that are responsible for adverse effects and for drug inactivation. Et3PAuCl was encapsulated in biocompatible PLGA–PEG nanoparticles (NPs) and the new formulation evaluated in colorectal HCT-116 cancer cells in comparison to the free gold complex. Notably, encapsulated Et3PAuCl (nano-Et3PAuCl hereafter) mostly retains the cellular properties of the free gold complex and elicits even greater cytotoxic effects in colorectal cancer (CRC) cells, mediated by apoptosis and autophagy. Moreover, a remarkable inhibition of two crucial signaling pathways, i.e. ERK and AKT, by nano-Et3PAuCl, was clearly documented. The implications of these findings are discussed.

The aim of this work is to identify the cisplatin binding sites on human H-chain ferritin. High-resolution X-ray crystallography reveals that cisplatin binds four distinct protein sites, that is, the side chains of His136 and Lys68, the side chain of His105, the side chain of Cys90 and the side chain of Cys102. These Pt binding sites are compared with those observed for the adduct that cisplatin forms upon encapsulation within horse spleen L-chain ferritin (87% identity with human L-chain ferritin).

A group of triethylphosphine gold(I) and silver(I) complexes, structurally related to auranofin, were prepared and investigated as potential anticancer drug candidates. The antiproliferative properties of these metal compounds were assessed against two leukemia cell lines, i.e., CCRF-CEM and its multidrug-resistant counterpart, CEM/ADR5000. Interestingly, potent cytotoxic effects were disclosed for both series of compounds against leukemia cells, with IC50 values generally falling in the low-micromolar range, the gold derivatives being on the whole more effective than the silver analogues. Some initial structure-function relationships were drawn. Subsequently, the ability of the study compounds to inhibit the three main catalytic activities of the proteasome was investigated. Different patterns of enzyme inhibition emerged for the various metal complexes. Notably, gold compounds were able to inhibit effectively both the trypsin-like and chymotrypsin-like proteasome activities, being less effective toward the caspase-like catalytic activity. In most cases, a significant selectivity of the study compounds toward the proteasome proteolytic activities was detected when compared to other proteases. The implications of the obtained results are discussed.

On the basis of two randomized trials, evolocumab and alirocumab have been approved in patients with cardiovascular disease. The evidence on these two agents has been studied through different methods of analysis that span from narrative approaches to network meta-analysis. In the present study, we assessed the performance of a narrative approach combined with the application of the restricted mean survival time (RMST). We studied the two pivotal placebo-controlled trials focused on evolocumab and alirocumab. Our original framework of comparative assessment employed the RMST. Our objective was to show that in the context of a narrative review, the RMST can be an efficient although simple tool to make indirect comparisons. The endpoint was event-free survival, expressed in months. For each cohort of patients (13,784 patients administered evolocumab, 9462 patients administered alirocumab, 23,242 controls), we determined the RMST values with 95% confidence intervals (CI) [evolocumab: 33.60 months, 95% CI 33.46–33.74; alirocumab: 34.07 months, 95% CI 33.92–34.22]. These results, along with those of the control groups, were analyzed and interpreted narratively. Univariate statistics were conducted, but no network meta-analysis was performed. The experience presented herein indicates that a framework of evidence assessment focused on the RMST is a worthwhile option. Our study is in line with the growing literature that has recently emphasized the methodological advantages of the RMST.

Dear Editor,Treatment-free remission (TFR) after discontinuation of a tyrosine-kinase inhibitor (TKI) has become an emerging goal for chronic myeloid leukaemia (CML) patients who have achieved a de...

Electrospray ionization mass spectrometry (ESI MS) is a powerful investigative tool to analyze the reactions of metallodrugs with proteins and peptides and characterize the resulting adducts. Here, we have applied this type of approach to four experimental anticancer gold(III) compounds for which extensive biological and mechanistic data had previously been gathered, namely, Auoxo6, Au2phen, AuL12, and Aubipyc. These gold(III) compounds were reacted with two representative proteins, i.e., human serum albumin (HSA) and human carbonic anhydrase I (hCA I), and with the C-terminal dodecapeptide of thioredoxin reductase. ESI MS analysis allowed us to elucidate the nature of the resulting metal-protein adducts from which the main features of the occurring metallodrug-protein reactions can be inferred. In selected cases, MS data were integrated and supported by independent 1HNMR and UV-Vis absorption measurements to gain an overall description of the occurring processes. From data analysis, it emerges that most of the investigated gold(III) complexes, endowed with an appreciable oxidizing character, undergo quite facile reduction to gold(I); the resulting gold(I) species tightly associate with the above proteins/peptides with a remarkable selectivity for free cysteine residues. In contrast, in the case of the less-oxidizing Aubipyc complex, the gold(III) oxidation state is conserved, and a gold(III) fragment still containing the original ligand is found to be associated with the target proteins. It is notable that the C-terminal dodecapeptide of thioredoxin reductase containing the characteristic -Gly-Cys-Sec-Gly metal-binding motif is able in all cases to trigger gold(III)-to-gold(I) reduction. Our investigation allowed us to identify in detail the nature of the gold fragments that ultimately bind the protein targets and determine the exact binding stoichiometry; some insight on the reaction kinetics was also gained. Notably, a few clear correlations could be established between the structure of the metal complexes and the nature of the resulting protein adducts. The mechanistic implications of these findings are analyzed and thoroughly discussed. Overall, the present results set the stage to better understand the real target biomolecules of these gold compounds and elucidate at the atomic level their interaction modes with proteins and peptides.

The clinically established gold drug Auranofin was reacted individually with a group of representative proteins, namely ubiquitin, ribonuclease A, carbonic anhydrase, haemoglobin and superoxide dismutase, and adduct formation was monitored in the various cases by ESI-MS analysis. We found that the reaction is highly selective for solvent exposed free cysteines that are modified through coordination of the AuPEt3+ fragment. Indeed, ESI-Q-TOF MS spectra carried out on protein samples incubated with a three fold molar excess of Auranofin allowed direct detection of the native proteins bearing bound AuPEt3+ fragments in the cases of carbonic anhydrase and haemoglobin. At variance, the two proteins that do not possess any free cysteine residue, i.e. ubiquitin and ribonuclease A, were unable to bind the gold fragment. In the case of superoxide dismutase, adduct formation is hindered by the scarce solvent accessibility of the free cysteine residue. These findings were further confirmed by a series of competition binding experiments with ebselen, a potent and selective cysteine-modifying reagent; we observed that pre-treatment with ebselen prevents the binding of the AuPEt3+ fragment to both carbonic anhydrase and haemoglobin.